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RESEARCH ARTICLE Z. Khiati, A.A. Othman & B.

Guessas, 20
S. Afr. J. Chem., 2007, 60, 20–24,
<http://journals.sabinet.co.za/sajchem/>.

Synthesis and Antibacterial Activity of 1,3,4-Oxadiazole and


1,2,4-Triazole Derivatives of Salicylic Acid and its
Synthetic Intermediates
Zoulikha Khiati , Adil A. Othman * and Bettache Guessas
1 1 2

1
Department of Chemistry, Faculty of Sciences, University of Sciences and Technology of Oran-Mohamed Boudiaf – USTO-MB,
P.O. Box 1505, El Mnaouer, Oran 31000, Algeria.
2
Laboratory of Applied Microbiology, Faculty of Sciences, University of Oran Es Senia, Oran 31000, Algeria.

Received 19 January 2006; revised 20 February 2007; accepted 5 March 2007.

ABSTRACT
Eight compounds 2–9 have been synthesized starting from salicylic acid, two of them (7 and 9) are novel. The four final products
namely: 5-(2-hydroxy phenyl)-1,3,4-oxadiazole-2-thione 4, 3-(2-hydroxy phenyl)-1H-1,2,4-triazole-5-thiol 6, 3-(2-hydroxy
phenyl)-4-amino-1,2,4-triazole-5-thiol 8 and 3-(2-hydroxy phenyl)-1-amino-1,2,4-triazole-5-thiol 9 have been prepared using
known reactions. The structures of intermediates and final products were determined by spectroscopic IR, UV, 1H-NMR &
MS-methods in addition to elemental analysis. Antibacterial activities of compounds 1–6 and 8 were investigated in vitro against
Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa and the results are reported herein.
KEYWORDS
1,3,4-Oxadiazole-2-thione; 1H-1,2,4-triazole-5-thiol; 4-amino-1,2,4-triazole-5-thiol; salicylic acid; antibacterial activity.

1. Introduction characteristic IR bands at 3400 cm–1 for OH and 3265 cm–1 for NH
Salicylic acid is widely used as antirheumatic and anti- and 1631 cm–1 for CO-N stretching.
inflammatory agents1 while compounds with the thiourea For the preparation of the oxadiazole 4, the hydrazide 3 was
–NH(CS)NH- function show antibacterial and antiviral activi- heated with an alcoholic solution of KOH and CS2 under reflux
ties.2 The 1,3,4-oxadiazoles and 1,2,4-triazoles are known to have conditions followed by acidification with HCl to give a brownish
a wide scope of biological and industrial activities. Among the crystalline product 4 in a very good yield (95 %). The mass
biological applications reported for 1,3,4-oxadiazole derivatives spectrum showed a molecular ion at 194.2 (M+) and the elemen-
are muscle relaxant,3 analgesic,3 hypnotic,3 sedative,3 CNS tal analysis corresponded with structure 4. The IR spectrum
depressing,4 tranquilizing,4 anticancer4 and antituberculostatic.5 showed an absorption at 1676 cm–1 for C=N which suggested
Furthermore the 1,3,4-oxadiazole derivatives have some indus- that compound 4 existed as the thione tautomer 4a rather than
trial applications in the fields of dyes,6 photosensivity,3,6 electrical the thiol form 4b which normaly exhibits the C=N stretching at
materials6 and liquid crystals3,6. lower region, i.e. in about 1638 cm–1 due to maximum conjuga-
Similarly 1,2,4-triazole derivatives have considerable biologi- tion.12 Further support for 4a came from the 1H-NMR spectrum
cal (antibacterial,7 antifungal7 and antitumor8) activities and which exhibited upfield singlets at 13.4 ppm and 10.54 ppm each
some industrial uses in the fields of photography9 and corrosion integrated for one proton which was designated to the NH and
inhibitors.10 In this paper we report the synthesis of different OH protons9,13 while the SH proton which is normally present at
heterocycles with the salicylic acid moiety represented by 3.5–6.5 ppm14 was absent. Other signals associated with the
compounds 4, 6, 8 and 9. aromatic protons appeared to match the expected signals (see
The thermal rearrangement between 8 and 9 is discussed the experimental section).
below. The antibacterial activities of the starting materials, the The synthesis of the triazole 6 was achieved by two steps from
synthetic intermediates and the products were tested and are the hydrazide 3, first by treatment of 3 with ammonium
reported below. thiocyanate and HCl for 15 hours under reflux conditions to give
the thiosemicarbazide derivative 5 as a crystalline product in a
2. Results and Discussion very good yield (94 %). The IR spectrum in CCl4 solution showed
a broad absorption in the region 3300–3100 cm–1 due to free and
2.1. Synthesis bonded OH and NH. The peak at 1660 cm–1 was assigned to
The final products 4, 6, 8 and 9 have been synthesized by a CO-N and the peak at 1241 cm–1 was assigned to C=S.14,15 The
common pathway as summarized in Scheme 1. The methyl mass spectrum showed the molecular ion fragments at m/z 138
salicylate 2 was synthesized in 90 % yield, and the IR spectrum for salicylamide, 120 for 2-hydroxy benzoyl and 91 for
showed an absorption at 1678 cm–1 for the CO-ester group which N-aminothiourea. The second step was achieved by heating 5 in
is in accordance with the literature.11 ethanolic KOH under reflux conditions followed by removal
The hydrazide 3 which is used as the starting material for the of the ethanol by vacuum distillation. A solid product was
common synthesis was obtained in 90 % yield by heating the extracted with ethyl acetate from the excess aqueous KOH layer.
ester 2 with hydrazine hydrate 64 %. The product exhibited The extract yielded the triazole 6 in 79 % after evaporation of the
* To whom correspondence should be addressed: E-mail: adilaliothman@yahoo.ca organic solvent. The IR spectra in THF solution showed charac-
RESEARCH ARTICLE Z. Khiati, A.A. Othman & B. Guessas, 21
S. Afr. J. Chem., 2007, 60, 20–24,
<http://journals.sabinet.co.za/sajchem/>.

Scheme 1
Synthesis of different heterocycles with the salicylic acid moiety.

teristic absorptions at 3300–3200 cm–1, 2861–2780 cm–1 for SH and Neither of these authors gave any physical evidence for the
1671 cm–1 for C=N although some tautomerism to thione might formation of substance 10. The infrared spectrum of the product
have taken place.13 we observed (7) showed characteristic absorption bands at
The 1H–NMR spectrum showed signals at 12.95 ppm and 3465 cm–1 (broad) for free and bonded N-H, at 1648 cm–1 for
10.05 ppm for NH and OH, respectively. CO-N and at 1447 cm–1 for C=S. The mass spectrum showed a
4-Amino triazole 8 was obtained by heating the oxadiazole 4 molecular ion (M+) at m/z 285.3 and a fragmentation ion at m/z
and hydrazine hydrate under reflux conditions for 8 h. The 201 which relates to 2-K+ –O-C6H4-CONHNHCS+. The elemen-
product formed crystalline fibres and had a slightly higher tal analysis of this compound correlated well with the formula
melting point 144 °C. The 1H–NMR spectrum showed signals at C8H6N2O3SK2 (see the experimental section).
13.2 ppm (NH2), 10.05 ppm (OH) and signals belonging to the Treatment of compound 7 with hydrazine hydrate is expected
phenylene protons at 8.04, 7.25, 6.9, 6.8 and 3.6 ppm for SH. The to give 9, the product was a crystalline solid which showed a
IR, MS spectrum and elemental analysis were in accordance lower melting point (87–88 °C) than the 4-amino-triazole (8)
with the structure for 8. which was prepared from the oxadiazole 4 as described above
The same compound 8 was also prepared according to the (Scheme 1). The IR, UV and elemental analysis for the product 9
method by Dimova2 and Zhang16. This involved the treatment of were similar to that of 8, but some differences were observed in
the hydrazide–3 with KOH/CS2 in ethanol and they reported the the 1H NMR spectrum. The NH signal showed at 13.8 , a signal at
formation of the dithio carbazinic acid–10. 12.25 for OH and the phenylene protons at 7.75, 7.45, 6.95,
O S 6.85 ppm were observed. On the basis of the mass spectrum
Cs2, KOH, EtOH which showed a molecular ion (M+) at m/z 208.1 and other
3 2 HO C6H4CNHNHCS-K+
spectral and elemental analysis data the structure of 1-amino-
10 triazole 9 was proposed. Formation of compound 9 might have
However, in our hands we isolated the novel 2N-(2-potassium resulted from the thermal rearrangement of the NH2 group in
oxy benzoyl)-potassium thiocarbazinic acid–7 (2-K+ –O-C6H4 position N-4 of 8 to position N-1 of 9, as a similar phenomenon
CONHNHCSO– K+) as a brownish crystalline with melting was reported before in the literature17,18. Molecular models of 9
point 196–197 °C in a yield of 68 %. suggest that the molecule prefers to exist in a coplanar form,
RESEARCH ARTICLE Z. Khiati, A.A. Othman & B. Guessas, 22
S. Afr. J. Chem., 2007, 60, 20–24,
<http://journals.sabinet.co.za/sajchem/>.

whereas 8 prefers the non-coplanar form. These differences are Table 1 Inhibition zones in (mm).
obviously affecting the polarity and the π electron distribution
between the two forms 8 and 9 and hence affecting their melting Compound* E. coli S. aureus P. aeruginosa
points and the NMR spectra.
Ampicellin 10 10 10
2.2. Biological Tests Gentamycin 10 08 12
1 6 7.5 6
The filter paper disk method (NCCLS)19,20 was employed in
2 9.5 6.5 0
duplicate for the in vitro study of antibacterial effects against the 3 6 0 0
Gram-negative bacteria Escherichia coli and Pseudomonas aerugi- 4a 8.5 6 0
nosa and Gram-positive bacteria Staphylococcus aureus using 5 8 6 0
Ampicellin and Gentamycin as references. The inhibitory effects 6 9 9 9
of compounds 1–6 and 8 against these bacteria are summarized 8 7 7.5 0
in Table 1 and are shown in Figs 1–3.
The screening results indicate that all the examined compounds, * Concentration 10 mg mL–1.
Key to the inhibition zones activities.
generally exhibit moderate activities against Gram-negative
Highly active = inhibition zone >12 mm.
bacteria E. coli and Gram-positive bacteria S. aureus as compared Moderately active = inhibition zone 9–12 mm.
with Ampicellin and Gentamycin. The triazole 6 has a better Slightly active = inhibition zone 6–9 mm.
inhibition effect than the Gentamycin against S. aureus since the Inactive = inhibition zone <6 mm.
latter is known to show some resistance to Gentamycin.21,22 On
the other hand the Gram-negative bacteria P. aeruginosa was only Microorganisms in this study were supplied by the university
affected by the triazole 6 and the salicylic acid 1. hospital of Oran and identified in the laboratory of applied
The inhibitory activities of the tested compounds on the microbiology, University of Oran Es Senia. The Mueller Hinton
Gram-negative and Gram-positive bacteria are arranged from medium was supplied by Difco.
higher to lower activities as follows.
Compounds 2 and 6 have a moderately active effect on the 3.2. Synthesis of Compounds
Gram-negative bacteria E. coli, while the compounds 5, 8 and 3 as
well as 1 exhibited slight activity against the same bacteria. Methyl-2-hydroxybenzoate 2 (Methylsalicylate)
Compounds 6, 8, 1, 2, 4a and 5 showed moderate to slight To a mixture of salicylic acid 1 (20.7 g, 0.15 mole) in methanol
activity against the Gram-positive bacteria S. aureus. (90 mL), conc., H2SO4 (16 mL) was added dropwise with stirring.
Compounds 4a and 1 showed moderate to slight activity The mixture was refluxed on a water bath at 80 °C for 5 h.
against the Gram-negative bacteria P. aeruginosa, while the TLC eluted with ethanol/benzene 1:4 showed Rf = 0.55 for the
compounds 2, 3, 4a, 5 and 8 exhibited no effect against the same acid 1 and Rf = 0.74 for the ester 2. Ice water (100 mL) was added
bacteria. at the end of the reaction with stirring. The aqueous mixture was
To confirm the above test, the minimum inhibition concentra- extracted two times with n-hexane (25 mL). The combined
tions were determined in liquid medium for the active organic layers were washed with 5 % aqueous NaHCO3 (150 mL)
substances for three times and the averages are shown in Table 2. until the pH reached 7 and then washed with 50 mL of water.
The organic layer was dried over anhydrous Na2SO4 and
3. Experimental filtered. The filtrate was evaporated to dryness to give a colour-
less oil; methyl salicylate 2 (20.6 g, yield 90 %); νmax/ cm–1 (liquid
3.1. General film) 3187 (OH), 1678 (CO). Lit 3190 (OH)11, 1675 (CO)11. UV
The melting points were measured with a BÜCHI 540 melting (methanol), λmax 235 nm. Lg ε 3.52. Lit. λmax 237. Lg ε 4.011.
point apparatus and are uncorrected. The IR spectra were
recorded using KBr discs and a JASCO V-530 spectrophotometer Salicylic Hydrazide 3
and in the IR spectra solutions were obtained with a GENESIS II Methylsalicylate 2 (5.0 g, 0.032 mole), ethanol (20 mL) and
FTIR spectrophotometer. The UV spectra were recorded on a hydrazine hydrate 64 % (6 mL) were mixed together and heated
ZUZI Split-Beam UV-Vis 4418PC (4418SPC) spectrophotometer. under reflux at 80 °C for 8 h. TLC eluted with ethanol/benzene
The 1H NMR (250 MHz) spectra were recorded in DMSO-d6 and 1:4 showed the development of a new spot at Rf = 0.51. Ethanol
the Mass spectra were recorded on a MAT 312 mass spectrometer was evaporated under reduced pressure and a white solid
using glycerol as matrix. product was recrystallized from H2O/MeOH to give salicylic

(1) (2) (3)

Figures 1–3 Antibiogrammes of (1) E.scherichia coli., (2) Staphylococcus aureus, and (3) Pseudomonas aeruginosa.
RESEARCH ARTICLE Z. Khiati, A.A. Othman & B. Guessas, 23
S. Afr. J. Chem., 2007, 60, 20–24,
<http://journals.sabinet.co.za/sajchem/>.

Table 2 Inhibition of microorganisms by compounds 1–6 and 8 at different concentrations.

Compound: 1 2 3 4a 5 6 8
Concentration‡ Concentration‡ Concentration‡ Concentration‡ Concentration‡ Concentration‡ Concentration‡
µg mL–1 µg mL–1 µg mL–1 µg mL–1 µg mL–1 µg mL–1 µg mL–1
a b c a b c a b c a b c a b c a b c a b c

i† + – – + + + + – – + + + + + + + + + + – –
ii + – – – – – – – – – – – – – – + + + – – –
iii + + – + – – – – – + – – + – – + + + + + –


Microorganisms: i, Escherichia coli; ii, Pseudomonas aeruginosa; iii, Staphylococcus aureus.

Concentrations (µg mL–1): a = 640 ; b = 320 ; c= 160.
Notes: the sign (+) for microorganism inhibitors and (–) for microorganisms intact.

hydrazide 3 (4.5 g, yield 90 %), m.p. 147 °C, lit. 147–150 °C23, and NH), 2861–2780 (SH) and 1671 (C=N); U.V. (methanol) λmax
νmax/ cm–1 (KBr) 3400 (OH), 3265 (NH), 1591 (CO-N), 1364 (Aro- 235, 300. Lg ε 3.55, 3.49, respectively. δH (250 MHz, DMSO-d6)
matic); UV (methanol) λmax 240, 300 nm. Lg ε 3.66, 3.55, respec- 12.95 (1H, s),10.05(1H,s), 7.75 (1H, d), 7.42 (1H, t), 6.95 (1H, d), 6.9
tively. (1H, t),4.2(1H,s). MS, 191.8 M+. (Found: C, 49.32; H, 03.52; N,
21.51 %. Calc. for C8H7N3OS (193.23); C, 49.74; H, 03.63; N,
5-(2-Hydroxyphenyl)-1,3,4-oxadiazole-2-thione 4a 21.76 %).
Salicylic hydrazide 3 (1.5 g., 0.01 mole) and ethanol (200 mL)
were added to a solution of KOH (0.84 g, 0.015 mole) in ethanol 2N-(2-Potassium oxybenzyl)-potassium Thiocarbazinic Acid 7
(20 mL) and CS2 (20 mL). The reaction mixture was heated under A mixture of salicylic hydrazide 3 (1.5 g, 0.01 mole) in ethanol
reflux at 80 °C for 9 h. The mixture became orange in colour. TLC 230 mL, alcoholic solution of KOH (8.4 g, 0.15 mole) in ethanol
eluted with ethanol/benzene 2:4 showed a product as Rf = 0.3. 15 mL and CS2 ( 9 mL 0.15 mol) were added dropwise and heated
Excess ethanol was removed under vacuum and the remainder under reflux on a water bath at 80 °C for 10 h. The ethanol was
of the solution was acidified with dil. HCl (10 %) to pH 5. A partially evaporated to 100 mL. The reaction mixture was cooled
brownish solid was filtered off and washed with ethyl acetate to room temperature, ether (200 mL) was added and a brownish
to dissolve the organic product. The washing solution upon precipitate was formed. The product was filtered off and
standing overnight at room temperature gave brown fibres washed twice with ether (50 mL), dried at room temperature to
which were recrystallized from CHCl3/EtOH to give 4 (1.85g, give a solid mass which was dissolved partially in warm
95 % yield), m.p. 167–168 °C; νmax / cm–1 (CCl4) 1676 (C=N), 1304– ethylacetate and filtered off. The filtrate was evaporated down
1251, 1157 (C-O-C); U.V. (methanol) λmax 235, 290, 300 nm. Lg ε to dryness to give brownish crystalline 7, recrystallized from
3.62, 3.54, 3.57, respectively; δH (250 MHz, DMSO-d6) 13.4(1H,s), chloroform/ethanol (1/1) to give the pure product (1.5 g, yield
10.54 (1H, s), 7.61 (1H, d), 7.31 (1H t), 6.97 (1H d), 6.92 (1H, t); MS, 59 %), m.p. 196–197 °C. νmax /cm–1 (THF) 3465 (NH), 1648 (CON);
194.2 M+. (Found: C, 49.21; H, 02.98; N, 14.39 %. Calc. for 1238 (C=S); U.V, λmax (methanol), 300, 290, 245 nm, Lg ε, 3.53, 3.50,
C8H6N2O2S (194.21); C, 49,48; H, 03.09; N, 14.43 %. 3.54, respectively. MS, 201 (2-KO-C6H4-CONHNHCS) and 285.3
(7); (Found: C, 37.51, H, 2.92, N, 09.26 %. Calc. for C8H6N2O3SK2
Thiosemicarbazide Salicylic Acid 5 (248); C, 33.33; H, 02.08; N, 09.7 %).
Salicylic hydrazide 3 (1g, 0.066 mole) was dissolved in ethanol
with stirring. Ammonium thiocyanate (1.6 g, 0.021 mole) and 3-(2-Hydroxy phenyl)-4-amino-1H-1,2,4-triazole-5-thiol 8
HCl (26 mL, 31 %) were added and the reaction mixture was The oxadiazole 4 (0.97 g, 0.005 mole) was dissolved in 80 mL
heated under reflux on a water bath for 15 h. TLC eluted with ethanol and hydrazine hydrate 64 % (10 mL) was added and the
ethanol/benzene 2:4 showed the development of a new spot, reaction mixture was heated under reflux on an water bath at
Rf = 0.55. Excess solvent was evaporated to almost dryness and 90 °C for 8 h. TLC (ethanol/benzene 2:4) gave a spot at Rf = 0.4.
the crystalline solid was filtered off and recrystallized from Excess ethanol was evaporated and the remaining solid/liquid
toluene/petroleum-ether 60–80 to give 5 (1.3 g, yield 94 %), m.p. mixture was filtered off and washed with ethyl acetate to give
144 °C; νmax / cm–1 (CCl4) 3300–3100 (br. OH, NH, stretching), 1660 yellowish-brown fibres (8) which was recrystallized from chlo-
(CO-N), 1610 (C=C), 1241 (C=S); U.V.(methanol) λmax 235, roform/ethanol (2/1) to give yellowish-brown crystals (0.75 g,
300 nm, Lg ε 3.49, 3.41. δ H (250 MHz, DMSO-d6) 13.8 yield 72 %), m.p. 144 °C; νmax / cm–1 (THF) 3500–3400 (OH, NH);
(4H,m),10.05(1H,s), 7.7 (1H, d), 7.4 (1H, t), 6.9 (1H, d), 6.8 (1H, t). 2681 (SH); UV.(methanol) λmax, 240, 285 nm, Lg ε 3.5, 3.30. δH
MS, 138 M + (salicylamide), 120 M + (2-oxy-benzoyl), 91 (250 MHz, DMSO-d6) 13.2(2H,s), 10.05(1H,s), 8.04 (1H, d), 7.25
M+(N-aminothio urea) (Found: C, 45.12; H, 4.09; N, 19.70 %. (1H, t), 6.9 (1H, d), 6.8 (1H, t),4.2(1H,s). MS 208,3; (Found: C,
Calc. for C8H9N3O2S (211.24); C, 45.50; H, 4.27; N, 19.9 %. 45.82; H, 3.7; N, 26.63 %. Calc. for C8H8N4OS (208.24); C, 46.15; H,
3.84; N, 26.92 %).
5-(2-Hydroxyphenyl)-1H-1,2,4-triazole-3-thiol 6
Thiosemicarbazide salicylic acid 5 (2.1 g, 0.01 mol) was 3-(2-Hydroxy phenyl)-1-amino-1H-1,2,4-triazole-5-thiol 9
dissolved in ethanol (200 mL). An ethanolic solution of KOH Compound 7 (1.1 g, 0.004 mole) dissolved in water (8 mL) and
(0.85 g, KOH, 0.015 mol) 20 mL was added and heated under hydrazine hydrate 64 % (4 mL) were heated under reflux on an
reflux at 80 °C for 4 h, to give one TLC spot (ethanol/benzene 2:4) oil bath at 110 °C for 6 h. TLC (ethanol/benzene 2:4) gave a spot at
at Rf = 0.11. Excess ethanol was evaporated to dryness and the Rf = 0.31. The reaction mixture was cooled to room temperature,
bulk of the solid was dissolved in ethyl acetate, filtered and iced-water 100 mL was added and the solution was made acidic
evaporated to dryness to give a solid which was recrystallized with 10 % HCl. A precipitate formed and was filtered off. The
from petroleum-ether/CH2Cl2 (1/3) to give brownish fibres 6 filtrate was extracted three times with 30 mL of ethyl acetate. The
(1.5 g, yield 79 %), m.p. 135 °C; νmax / cm–1 (THF) 3300–3200 (OH extracts were combined and dried over anhydrous Na2SO4. The
RESEARCH ARTICLE Z. Khiati, A.A. Othman & B. Guessas, 24
S. Afr. J. Chem., 2007, 60, 20–24,
<http://journals.sabinet.co.za/sajchem/>.

filtrate was evaporated to dryness to give a brown solid 9, which 4 A. Mohsen, M. Omar and D. Aboul Wafa, J. Heterocyclic. Chem., 21,
was recrystallized from chloroform/ethanol (2/1) to give of 1984, 1415–1418.
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3500–3400 (OH, NH), 2677 (SH); U.V. (methanol) λmax, 230 nm.
6 Y. Zhang, R-Z. Qiao, P-F. Xu, Z-Y. Zhang, Q. Wang, L-M. Mao and K-B.
Lg ε 3.48. δH (250 MHz, DMSO-d6) 13.85(2H,s), 10.25 (2H, s), 7.75 Yu, J. Chin. Chem. Soc., 2002, 49, 369–373.
(1H, t), 7.45 (1H, t), 6.95 (1H, d), 6.85 (1H, t), 3.9(1H,s). MS, 208.1 7 L. Jianbing, L. Lichun, D. Hong, L. Zhun and F. Jianxin, J. Organometal.
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3.3. Antibacterial Tests 9 A. Shawali, I. Zeid, M. Abdelkader, A. Elsherkini and F. Altalbawy,
The filter paper disc method was performed in duplicate using J. Chin. Chem. Soc., 2001, 48, 65–72.
fresh Mueller Hinton agar medium. This agar medium was 10 L.J. Berchmans, V. Sivan and S.V.K. Iyer, Mat. Chem. and Phys., 2006,
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inoculated with 0.5 mL of cultures containing about 106 CFU/mL.
11 M. Chavanne, A. Jullien, G.J. Beaudoin and E. Flamand, Chimie
Filter paper discs (5 mm diameter) saturated with solutions of organique expérimentale, 2ème édition, Modulo Editeur, 1991, p. 574.
each compound (concentrations 10 mg mL–1 ethanol) was placed 12 F. Aydogan, Z. Turgut and N. Ocal, Turk. J. Chem., 2002, 26, 159–169.
on the indicated agar mediums. The incubation time was 24 h 13 A. Zhang, L. Zhang and X. Lei, Magn. Reson. Chem., 2006, 44, 813–816.
at 37 °C. The blank test disc with ethanol was used. Inhibitory
14 K. Zamani, K. Faghihi, M.R. Sangi and J. Zolgharnein, Turk. J. Chem.,
activity was evaluated by measuring the diameter of clear zone 2003, 27, 119–125.
observed around the disc (in mm). 15 A. Shawali, M. Abdallah, M. Mosselhi and Y. Mohamed, Z. Natur-
forsch., 2002, 57b, 552–556.
Acknowledgements 16 L. Zhang, A. Zhang, X. Chen, X. Lei, X. Nan, D. Chen and Z. Zhang,
Thanks are due to Prof. N. Al-Masoudi, Department Chemie, Molecules, 2002, 7, 681–689.
Universität Konstanz, Konstanz, Germany, for 1H-NMR, MS and 17 P. Carisen, O.R. Gauton and K.R. Jorgensen, Molecules, 1996, 1,
elemental analysis, Prof. M. Kihal, Uni-Oran, for useful discus- 242–250.
sion, and Mrs H. Boubekeur and Miss I. Abboub for technical 18 K.B. Jorgensen, R.B. Olsen and P.H.J. Carrisen, Molecules, 2001, 6,
481–195.
assistance.
19 National Committee for Clinical and Laboratory Standards
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